阴道毛滴虫黏附蛋白33基因原核表达载体的构建及表达  

Construction and Expression in vitro of Prokaryotic Expressional Vector of Trichomonas vaginalis Adhesion Protein 33 Gene

作　　者：杨树国[1] 王雅静[2] 朱晓燕[2] 帖超男[2] 廖琳[2] 

机构地区：[1]郧阳医学院寄生虫学教研室,湖北十堰442000 [2]四川大学华西基础医学与法医学院寄生虫学教研室

出　　处：《四川动物》2008年第6期1021-1023,共3页Sichuan Journal of Zoology

摘　　要：目的构建阴道毛滴虫黏附蛋白33基因原核表达载体并诱导其体外表达。方法pMD-18T-ap33重组质粒和pUC18空质粒经BamH Ⅰ和XbaⅠ限制性内切酶双酶切,将ap33基因亚克隆入pUC18载体并进行筛选和鉴定。重组质粒经IPTG诱导,SDS-PAGE电泳及Western-blot杂交鉴定重组蛋白。结果经双酶切及PCR鉴定,构建的重组质粒为阳性重组子,诱导出的重组蛋白大小约为Mr36000,与理论值基本相符。结论成功构建重组质粒并获得体外表达。Objective To construct the prokaryotic expressional vector of adhesion protein 33 （ap33） on the isolate of Trichomonas vaginalis and induce the expression of ap33 gene in vitro. Methods The plasmid pMD-18T-ap33 and pUC18 were digested by BamH Ⅰ and Xba Ⅰ . The ap33 gene was subeloned into the plasmid pUC18 and induced to express AP33 in prokaryotic cell by IFTG, which was analyzed by SDS-PAGE and Western-blot. Results The correct recombinant plas- mid pUC18-ap33 was isolated and confirmed by PCR and restriction analysis. The relative molecular mass of AP33 was almost Mr 36 000. Conclusion The recombinant plasmid pUC18-ap33 was constructed sueeessfully and the adhesion protein 33 could be induced to express in vitro.

关 键 词：阴道毛滴虫 黏附蛋白33 原核表达载体 基因表达 

分 类 号：Q78[生物学—分子生物学]